The evaluation of expression of genes in gamma globin synthesis before and after differentiation of hematopoietic stem cells into erythroid lineage

Induction of fetal hemoglobin (Hb-F) can improve the patients’ symptoms of haemoglobinopathies. Several factors can induce gamma globin gene expression and increased Hb-F levels in patients. In this study, the expression of genes is involved in regulation of gamma globin synthesis such as PIPKII-alpha BCL11a, and miR-30a during CD34 hematopoietic stem cell differentiation into erythroid lineage was studied.
Material and In this study, CD34 cells were isolated from umbilical cord blood. Then, CD34 cells were cultured in differentiation medium. Erythroid colonies were harvested on days 8, 11, 14 and gene expression of gamma globin, PIPKII-alpha, BCL11a, and miR-30a were analyzed by quantitative RT- PCR.
The results showed that gene expression of gamma-globin gradually at the 8th day after differentiation started and at 14th day reached to the highest level of its expression. In parallel with increasing the expression of gamma-globin, the expression of PIPKII-alpha increased, while the expression of BCL11a decreased. Moreover, the expression of miR- 30a increased during differentiation into erythroid lineage.
The results indicated that increased expression of PIPKII-alpha and decreased expression of BCL11a during CD34 cells differentiation to erythroid lineage. According to expression pattern of miR-30a and its target genes, PIPKII-alpha and BCL11a, we cannot suppose the inhibitory effect of miR-30a on PIPKII-alpha expression, but due to inhibitory effect of miR-30a on BCL11a gene, we can introduce this microRNA as a regulatory molecule in the induction of Hb-F pathway. However, the further studies are needed to investigate this interaction.